論文 - 詳細
| RRC ID | 85170 |
|---|---|
| 著者 | Matsuyama A, Hashimoto A, Arioka M, Yoshida M. |
| タイトル | Improvement of Targeting Efficiency by Promoter Replacement of Markers in Integration Vectors. |
| ジャーナル | Genes Cells |
| Abstract |
To establish a gene expression system that reflects physiological conditions, we developed a series of vectors that can be integrated into the chromosome. Compared with the integration vectors employing double-crossover recombination, single-crossover integration vectors have the advantage of high transformation efficiency. However, because single-crossover recombination generates repeat sequences upstream and downstream of the integrated fragment, this strategy is often associated with a risk that an integrated fragment may pop out from the chromosome during cultivation. Here, we assessed the frequency of pop-out using a fission yeast single-crossover integration vector, pDUAL. We also examined the effect of shortening the repeats on pop-out by employing a strategy involving heterologous replacement of the promoter for the leu1 marker in the vector. Due to the intrinsic low frequency of pop-out, the effect of promoter conversion on pop-out was negligible, if any. However, a clear ameliorative effect was observed in obtaining the desirable transformants in which a vector fragment was correctly inserted at the targeted locus, a result that may be driven by the limited potential for recombination in the promoter replacement construct. |
| 巻・号 | 30(2) |
| ページ | e70013 |
| 公開日 | 2025-3-1 |
| DOI | 10.1111/gtc.70013 |
| PMID | 40110920 |
| MeSH | Gene Targeting* / methods Genetic Markers Genetic Vectors* / genetics Promoter Regions, Genetic* Recombination, Genetic Schizosaccharomyces* / genetics |
| IF | 1.655 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | X(Twitter) |
| 各媒体での言及数の合計 | 1 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| 酵母 | FY47720 FY47734 |